本文選題：熒光方法 + 支撐膜�。� 參考：《伊犁師范學院》2015年碩士論文

【摘要】：隨著納米技術(shù)在生物醫(yī)學方面的發(fā)展,控制藥物向固定的病灶接近,并在接近后進行藥物的緩釋,可以使藥物發(fā)揮出最大的療效,即提高藥物的靶向型,成為當前生物醫(yī)學研究領域的關(guān)注點。為了更有效的控制藥物做定向移動,并且定向向病灶做緩解,需要選擇合適的藥物載體以提高藥物的靶向性。由于金納米顆粒和脂質(zhì)體在做藥物載體方面表現(xiàn)出一些優(yōu)異的特性,因而研究金納米顆粒的吸附特性、利用滲透壓方式調(diào)整脂質(zhì)體藥物輸運的內(nèi)吞方式,成為當前生物醫(yī)學前沿領域的研究熱點。磷脂雙層膜在納米生物技術(shù)領域展現(xiàn)出廣闊的應用前景。傳感器基座、藥物載體等固體表面的磷脂膜自組織行為逐漸成為材料、物理與生物交叉學科的重要問題。本實驗研究基于磷脂支撐雙層膜體系,以石英電子微天平、熒光顯微鏡為主要實驗表征手段,研究了二氧化硅表面的磷脂膜融合、半融合、顆粒吸附自組織行為機理及其調(diào)控方法。一方面基于金納米顆粒對特定的熒光波段的淬滅效應,利用熒光顯微鏡、以及熒光能量共振轉(zhuǎn)移(FRET)技術(shù),在構(gòu)建正、倒置樣品實驗艙上,研究了不同尺寸金納米顆粒在仿生膜上吸附的重力產(chǎn)生的影響。另一方面,以石英晶體微天平與耗散系數(shù)測量儀、納米粒度儀等為實驗主要的表征手段,研究了高低滲透壓梯度對納米尺寸的磷脂囊泡在二氧化硅襯底表面的吸附和融合動力學過程的影響。本碩士論文第二章工作中,研究了重力在顆粒吸附過程中發(fā)揮的重要作用。納米顆粒在生物膜表面的吸附行為是納米生物技術(shù)領域的重要問題。本實驗研究采用熒光手段,建立正、倒置實驗模型,定量地建立了溶液頂層、底層支撐膜表面顆粒吸附量的差異量與顆粒的沉淀、擴散速率之間的關(guān)系,借助熒光顯微鏡觀察重力對金納米顆粒在支撐磷脂膜表面的吸附影響,并對不同顆粒尺寸在其頂層或底層支撐膜表面吸附量的差異性作了系統(tǒng)的觀察研究和理論分析。實驗研究發(fā)現(xiàn)：顆粒尺寸決定其在頂或底層支撐膜表面吸附的差異性.吸附量的差異與顆粒的沉淀速率和擴散速率之比的對數(shù)呈線性關(guān)系。當顆粒的直徑小于14 nm時,則無需考慮重力在顆粒吸附過程中的影響;當顆粒的直徑大于176 nm時,就發(fā)現(xiàn)重力在吸附過程中占主導的地位。這一研究為藥物載體、理解納米顆粒與生物膜之間的相互作用提供了參考依據(jù)。在理想情況下,磷脂囊泡在親水表面可自發(fā)融合形成支撐膜。本碩士論文第三章工作中,我們研究了不同滲透壓梯度對納米級囊泡在二氧化硅表面吸附、融合、破裂及向磷脂雙層膜過程轉(zhuǎn)變速率的影響。實驗開始時,用石英晶體微天平與耗散系數(shù)測量儀(QCM-D)來獲取在等滲環(huán)境下時間參數(shù)之間的定量關(guān)系。然后用石英晶體微天平與耗散系數(shù)測量儀(QCM-D)來研究時間參數(shù)的變化與高、低滲環(huán)境下囊泡融合速率之間的關(guān)系。最后用納米粒度儀表征在不同滲透條件下囊泡的形狀特征,分析了囊泡對稱性破壞和膜的表面張力對囊泡融合進程快慢影響的可能機理。實驗研究發(fā)現(xiàn),糖的存在對囊泡融合速率影響較小。在低滲滲透壓梯度下,在測試的整個滲透壓梯度范圍內(nèi),滲透壓作用力對囊泡融合過程的影響均很小；而在高滲滲透壓梯度下,當滲透壓梯度小于200m0sm時,此融合影響效應不明顯,當滲透壓梯度高于該值時,則對融合進程的快慢有顯著影響。能量分析說明,與滲透壓相關(guān)的化學勢能主要以跨磷脂膜定向水擴散的形式釋放。因此相對于表面粘滯力,滲透壓對吸附囊泡的形變與融合影響較小。我們的研究深化了滲透壓在膜融合過程中作功方式的理解,并為在仿生器件構(gòu)筑中合理地使用滲透壓提供指導。最后,我們對論文進行了總結(jié),并對今后工作進行了展望。
[Abstract]:With the development of nanotechnology in the biomedical field, the control of drugs to the fixed focus, and the release of the drug after approaching, can make the drug play the most effective, that is, to improve the target type of the drug, and become the focus of the current biomedical research field. To remission to the focus, it is necessary to choose the appropriate drug carrier to improve the targeting of the drug. Because gold nanoparticles and liposomes have shown some excellent properties in making drug carriers, the adsorption characteristics of gold nanoparticles are studied and the endocytosis method of liposome transport by osmotic pressure is adjusted to become the current biomedicine. The phospholipid bilayer film shows a broad application prospect in the field of nanoscale biotechnology. The self-organizing behavior of phospholipid membrane on solid surface of sensor base, drug carrier and other solid surfaces gradually becomes an important problem in the physical and biological interdisciplinary. This experiment is based on the phospholipid braced double layer membrane system with quartz electron Microbalance and fluorescence microscopy are the main means of experimental characterization. The mechanism and control methods of phospholipid membrane fusion, semi fusion and self-organizing particle adsorption on silica surface are studied. On the one hand, the quenching effect of gold nanoparticles on specific fluorescent bands, fluorescence microscopy, and fluorescence energy resonance transfer (FRET) technology are used. The effect of the gravitational effect of different sizes of gold nanoparticles on the biomimetic membrane was studied in a positive and inverted sample experiment module. On the other hand, the quartz crystal microbalance and dissipation factor measuring instrument and the nano particle size meter were used as the main characterization means to study the high and low osmosis pressure gradient on the nano size phospholipid vesicles in two oxygen. The influence of adsorption and fusion kinetics on the surface of silicon substrate. The important role of gravity in the process of particle adsorption is studied in the second chapter of the master's thesis. The adsorption behavior of nanoparticles on the surface of the biofilm is an important problem in the field of nanoscale biotechnology. The relationship between the difference of the amount of particle adsorption on the surface of the surface of the support membrane and the precipitation of the particles and the rate of diffusion is established quantitatively, and the effect of gravity on the adsorption of the gold nanoparticles on the surface of the phospholipid membrane is observed by the fluorescence microscope, and the amount of adsorption of different particle sizes on the top or bottom support membrane surface is also observed. The difference is made by systematic observation and theoretical analysis. The experimental study shows that the particle size determines the difference between the adsorption on the surface of the top or the bottom support membrane. The difference of the amount of adsorption is linear with the logarithm of the ratio of the precipitation rate to the diffusion rate. When the diameter of the particle is less than 14 nm, there is no need to consider the gravity in the particle suction. The influence of the process is attached. When the diameter of the particle is greater than 176 nm, it is found that gravity is dominant in the process of adsorption. This study provides a reference for the understanding of the interaction between nanoparticles and biofilms. In the ideal case, the phospholipid vesicles can be spontaneously fused into a supporting membrane on the surface of the water surface. In the third chapter, we studied the effects of different osmotic pressure gradients on the adsorption, fusion, rupture and transition rate of nanoscale vesicles on the silica surface. At the beginning of the experiment, the quantitative relationship between the time parameters of the isosotic environment was obtained by using quartz crystal microbalance and QCM-D. Then the quartz crystal microbalance and the dissipative coefficient measuring instrument (QCM-D) were used to study the relationship between the change of time parameters and the rate of vesicle fusion in the high and low permeability environment. Finally, the shape characteristics of the vesicles were characterized by the nano particle size instrument under different infiltration conditions, and the bubble symmetry destruction and the surface tension of the membrane to the vesicle fusion process were analyzed quickly. It is found that the presence of sugar has little effect on the rate of vesicle fusion. Under the low osmotic pressure gradient, the influence of osmotic pressure on the process of vesicle fusion is very small in the range of the whole osmotic pressure gradient in the test, and when the osmotic pressure gradient is less than 200m0sm The effect is not obvious. When the osmotic pressure gradient is higher than that, it has a significant effect on the speed of the fusion process. The energy analysis shows that the chemical potential energy related to osmotic pressure is mainly released in the form of directional water diffusion across the phospholipid membrane. Therefore, relative to the surface viscosity, the osmotic pressure has little effect on the deformation and fusion of the adsorbed vesicles. The study deepens the understanding of the work mode of osmotic pressure in the process of membrane fusion and provides guidance for the rational use of osmotic pressure in the construction of biomimetic devices. Finally, we summarize the paper and look forward to the future work.

【學位授予單位】：伊犁師范學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：TB383.1;O647.3

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